Detection of bacteria and fungi in blood of patients with febrile neutropenia by real-time PCR with universal primers and probes

Teranishi, Hideto; Ohzono, Nanae; Inamura, Norikazu; Kato, Atsushi; Wakabayashi, Tokio; Akaike, Hiroto; Terada, Kihei; Ouchi, Kazunobu

doi:10.1016/j.jiac.2014.11.008

Cited by 10 publications

(5 citation statements)

References 15 publications

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“…Similar limitations have been encountered by other authors using in-house or commercially available PCR-based assays (Teranishi et al 2015;Ziegler et al 2016), designed to detect infectious agents directly in blood samples.…”

Section: Rapid Methods For Detection Of Microorganisms Directly In Bl...mentioning

confidence: 55%

“…The possibility to rapidly detect and identify microbial pathogens directly in blood samples from patients with suspected sepsis is an attractive goal since it would allow to obtain precious diagnostic information in a few hours from blood drawing. Most of the proposed solutions involve the use of PCR assays specifically designed to detect microorganisms in blood samples (Mencacci et al 2012;Pasqualini et al 2012;Gosiewski et al 2014;Teranishi et al 2015;Ziegler et al 2016). To this aim, Gosiewski et al (2014) compared four in-house developed methods (nested PCR, multiplex PCR, qPCR, and FISH) and the SeptiFast assay with the BacT/ALERT 3D blood culture system (bioM erieux) for their ability to detect the presence of microbes in 71 blood samples of patients with clinical symptoms of sepsis.…”

Section: Rapid Methods For Detection Of Microorganisms Directly In Bl...mentioning

confidence: 99%

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Recent advances in the microbiological diagnosis of bloodstream infections

Florio

Morici

Ghelardi

et al. 2017

Critical Reviews in Microbiology

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Rapid identification (ID) and antimicrobial susceptibility testing (AST) of the causative agent(s) of bloodstream infections (BSIs) are essential for the prompt administration of an effective antimicrobial therapy, which can result in clinical and financial benefits. Immediately after blood sampling, empirical antimicrobial therapy, chosen on clinical and epidemiological data, is administered. When ID and AST results are available, the clinician decides whether to continue or streamline the antimicrobial therapy, based on the results of the in vitro antimicrobial susceptibility profile of the pathogen. The aim of the present study is to review and discuss the experimental data, advantages, and drawbacks of recently developed technological advances of culture-based and molecular methods for the diagnosis of BSI (including mass spectrometry, magnetic resonance, PCR-based methods, direct inoculation methods, and peptide nucleic acid fluorescence in situ hybridization), the understanding of which could provide new perspectives to improve and fasten the diagnosis and treatment of septic patients. Although blood culture remains the gold standard to diagnose BSIs, newly developed methods can significantly shorten the turnaround time of reliable microbial ID and AST, thus substantially improving the diagnostic yield.

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Section: Rapid Methods For Detection Of Microorganisms Directly In Bl...mentioning

confidence: 55%

Section: Rapid Methods For Detection Of Microorganisms Directly In Bl...mentioning

confidence: 99%

Recent advances in the microbiological diagnosis of bloodstream infections

Florio

Morici

Ghelardi

et al. 2017

Critical Reviews in Microbiology

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“…This finding inconsistent with previous observations, detecting the infecting microorganisms in only 20–30% of FN episodes. 21,22 Accordingly, this may partially explain the physicians’ deviation from guideline recommendations. The persistence of fever might have encouraged physicians to add or switch antibiotics empirically when the source of infection could not be found.…”

Section: Discussionmentioning

confidence: 99%

Adherence to practice guidelines for the management of febrile neutropenia in patients undergoing hematopoietic stem cell transplantation: An observational study in a referral center in Iran

Kolour

Shahrami

Kargar

et al. 2022

J Oncol Pharm Pract

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Introduction Patients undergoing hematopoietic stem cell transplantation (HSCT) are suspected to develop febrile neutropenia (FN) and severe infections. Therefore, appropriate prescription of antibiotics in these patients is crucial to reduce the rates of morbidity, mortality, and antimicrobial resistance. The present study aimed to evaluate the physicians’ prescription and adherence to the FN clinical guidelines among patients undergoing HSCT. Methods This prospective observational single-center study was conducted during a 15-month period in a tertiary referral hospital in Iran. The patients with at least one episode of FN following HSCT were included in the current study. The physicians’ adherence to the Infectious Diseases Society of America (IDSA) and National Comprehensive Cancer Network (NCCN) clinical guidelines for the management of FN was evaluated using prescription data and medical record reviews. Results Two hundred and fifteen patients with 297 FN episodes were evaluated. The timing of antibiotics and the selection of the initial regimen were considered guideline-based therapy. However, antibiotic dosing and initial regimen modification were not followed in terms of the guideline recommendations in 58.1% of the patients. In particular, vancomycin was inappropriately given in 83.1% of patients. The overall adherence of physicians to the guidelines was 35.8%. Conclusion Non-adherence to clinical guidelines is high particularly in initial regimen modification and administration of vancomycin, which affects hospital stay and patient's outcome. Implementation of guideline-review sessions to raise the awareness of the physicians and to improve the rational use of antimicrobial agents may be crucial.

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“…With the rapid development of molecular biology technology, PCR technology was considered to be a good choice in addition to the above techniques, and has been more commonly applied to pathogenic microorganism detection. Compared with traditional methods, PCR technology has the advantages of high specificity, sensitivity, and rapidity [26,27]. However, PCR requires expensive thermal cycling equipment, and finally requires agarose gel electrophoresis to further verify the detection results, which was time-consuming and inapplicable.…”

Section: Discussionmentioning

confidence: 99%

CelB is a suitable marker for rapid and specific identification of Klebsiella pneumoniae by the loop-mediated isothermal amplification (LAMP) assay

et al. 2019

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Klebsiella pneumoniae belongs to Enterobacteriaceae, which is the commonest bacterium causing nosocomial respiratory tract infection. It ranks second in bacteremia and urinary tract infection in gram-negative bacteria. Therefore, the rapid and accurate identification of K. pneumoniae was of great significance for the guide of clinical medication, and timely treatment of patients. The purpose of this study was to establish a rapid and sensitive molecular detection method for K. pneumoniae based on loopmediated isothermal amplification (LAMP) technology. Firstly, local BLAST and NCBI BLAST were used to analyze the genome of K. pneumoniae. According to the principle of interspecific and intraspecific specificity, CelB (GenBank ID 11847805) was selected as the specific gene. Then, the LAMP and PCR identification systems were established with this target gene. Thirty-six clinical isolates of K. pneumoniae and 50 non-K. pneumoniae were used for the specific evaluation, and both LAMP and PCR could specifically distinguish K. pneumoniae from non-K. pneumoniae. A 10-fold series diluted positive plasmids and simulated infected blood samples were used as the templates in the sensitivity assay, and the results showed that the sensitivity could reach 1 copy/reaction. In summary, a rapid, specific, and sensitive LAMP method was established to detect K. pneumoniae in clinics.

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Detection of bacteria and fungi in blood of patients with febrile neutropenia by real-time PCR with universal primers and probes

Cited by 10 publications

References 15 publications

Recent advances in the microbiological diagnosis of bloodstream infections

Recent advances in the microbiological diagnosis of bloodstream infections

Adherence to practice guidelines for the management of febrile neutropenia in patients undergoing hematopoietic stem cell transplantation: An observational study in a referral center in Iran

CelB is a suitable marker for rapid and specific identification of Klebsiella pneumoniae by the loop-mediated isothermal amplification (LAMP) assay

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